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IntroductionHigh-throughput phenotyping technologies together with metabolomics analysis can speed up the development of highly efficient and effective biostimulants for enhancing crop tolerance to drought stress. The aim of this study was to examine the morphophysiological and metabolic changes in tomato plants foliarly treated with two protein hydrolysates obtained by enzymatic hydrolysis of vegetal proteins from Malvaceae (PH1) or Fabaceae (PH2) in comparison with a control treatment, as well as to investigate the mechanisms involved in the enhancement of plant resistance to repeated drought stress cycles. MethodsA phenotyping device was used for daily monitoring morphophysiological traits while untargeted metabolomics analysis was carried out in leaves of the best performing treatment based on phenotypic results.ResultsPH1 treatment was the most effective in enhancing plant resistance to water stress due to the better recovery of digital biomass and 3D leaf area after each water stress event while PH2 was effective in mitigating water stress only during the recovery period after the first drought stress event. Metabolomics data indicated that PH1 modified primary metabolism by increasing the concentration of dipeptides and fatty acids in comparison with untreated control, as well as secondary metabolism by regulating several compounds like phenols. In contrast, hormones and compounds involved in detoxification or signal molecules against reactive oxygen species were downregulated in comparison with untreated control.ConclusionThe above findings demonstrated the advantages of a combined phenomics-metabolomics approach for elucidating the relationship between metabolic and morphophysiological changes associated with a biostimulant-mediated increase of crop resistance to repeated water stress events.

The demand for high-quality strawberries continues to grow, emphasizing the need for innovative agricultural practices to enhance both yield and fruit quality. In this context, the utilization of natural products, such as biostimulants, has emerged as a promising avenue for improving strawberry production while aligning with sustainable and eco-friendly agricultural approaches. This study explores the influence of a bacterial filtrate (BF), a vegetal-derived protein hydrolysate (PH), and a standard synthetic auxin (SA) on strawberry, investigating their effects on yield, fruit quality, mineral composition and metabolomics of leaves and fruits. Agronomic trial revealed that SA and BF significantly enhanced early fruit yield due to their positive influence on flowering and fruit set, while PH treatment favored a gradual and prolonged fruit set, associated with an increased shoot biomass and sustained production. Fruit quality analysis showed that PH-treated fruits exhibited an increase of firmness and soluble solids content, whereas SA-treated fruits displayed lower firmness and soluble solids content. The ionomic analysis of leaves and fruits indicated that all treatments provided sufficient nutrients, with heavy metals within regulatory limits. Metabolomics indicated that PH stimulated primary metabolites, while SA and BF directly affected flavonoid and anthocyanin biosynthesis, and PH increased fruit quality through enhanced production of beneficial metabolites. This research offers valuable insights for optimizing strawberry production and fruit quality by harnessing the potential of natural biostimulants as viable alternative to synthetic compounds.

IntroductionThe fermentative production of auxin/indole 3-acetate (IAA) using selected Pantoea agglomerans strains can be a promising approach to developing novel plant biostimulants for agriculture use.MethodsBy integrating metabolomics and fermentation technologies, this study aimed to define the optimal culture conditions to obtain auxin/IAA-enriched plant postbiotics using P. agglomerans strain C1. Metabolomics analysis allowed us to demonstrate that the production of a selected.Results and discussion:Array of compounds with plant growth-promoting- (IAA and hypoxanthine) and biocontrol activity (NS-5, cyclohexanone, homo-L-arginine, methyl hexadecenoic acid, and indole-3-carbinol) can be stimulated by cultivating this strain on minimal saline medium amended with sucrose as a carbon source. We applied a three-level-two-factor central composite design (CCD) based response surface methodology (RSM) to explore the impact of the independent variables (rotation speed and medium liquid-to-flask volume ratio) on the production of IAA and IAA precursors. The ANOVA component of the CCD indicated that all the process-independent variables investigated significantly impacted the auxin/IAA production by P. agglomerans strain C1. The optimum values of variables were a rotation speed of 180 rpm and a medium liquid-to-flask volume ratio of 1:10. Using the CCD-RSM method, we obtained a maximum indole auxin production of 208.3 ± 0.4 mg IAAequ/L, which was a 40% increase compared to the growth conditions used in previous studies. Targeted metabolomics allowed us to demonstrate that the IAA product selectivity and the accumulation of the IAA precursor indole-3-pyruvic acid were significantly affected by the increase in the rotation speed and the aeration efficiency.

Co-inoculation of arbuscular mycorrhizal fungi (AMF) and bacteria can synergically and potentially increase nitrogen use efficiency (NUE) in plants, thus, reducing nitrogen (N) fertilizers use and their environmental impact. However, limited research is available on AMF-bacteria interaction, and the definition of synergisms or antagonistic effects is unexplored. In this study, we adopted a response surface methodology (RSM) to assess the optimal combination of AMF (Rhizoglomus irregulare and Funneliformis mosseae) and Bacillus megaterium (a PGPR—plant growth promoting rhizobacteria) formulations to maximize agronomical and chemical parameters linked to N utilization in maize (Zea mays L.). The fitted mathematical models, and also 3D response surface and contour plots, allowed us to determine the optimal AMF and bacterial doses, which are approximately accorded to 2.1 kg ha–1 of both formulations. These levels provided the maximum values of SPAD, aspartate, and glutamate. On the contrary, agronomic parameters were not affected, except for the nitrogen harvest index (NHI), which was slightly affected (p-value of < 0.10) and indicated a higher N accumulation in grain following inoculation with 4.1 and 0.1 kg ha–1 of AMF and B. megaterium, respectively. Nonetheless, the identification of the saddle points for asparagine and the tendency to differently allocate N when AMF or PGPR were used alone, pointed out the complexity of microorganism interaction and suggests the need for further investigations aimed at unraveling the mechanisms underlying this symbiosis.

In this work, the impact of terroir, cultivar, seasonality, and farming systems on functional traits of tomato was hierarchically investigated. Untargeted metabolomics, antioxidant capacity, colorimetric assays, and enzyme inhibition were determined. The total phenolic and carotenoid contents significantly varied between growing years, whereas an interaction between the farming system and growing year (p < 0.01) was observed for total phenolics, carotenoids, and flavonoids, and for acetylcholinesterase inhibition. Hierarchical clustering showed that geographical origin and growing year were the major contributors to the differences in phytochemical profiles. Nonetheless, supervised modeling allowed highlighting the effect of the farming system. Several antioxidants (L-ascorbic acid, α-tocopherol, and 7,3′,4′-trihydroxyflavone) decreased, whereas the alkaloid emetine and phytoalexin phenolics increased under organic farming. Taken together, our findings indicate that cultivar and pedo-climatic conditions are the main determinants for the functional quality of tomato, whereas the farming system plays a detectable but hierarchically lower.

The application of microbial biostimulants to plants has revealed positive effects related to nutrients uptake, stress tolerance, root development and phenological growth. However, little information is available exploiting the potential synergistic biostimulant action of microbes on the functional quality of the yields. The current research elucidated the effect of single or coupled action of biostimulants, associated with either optimal or reduced nitrogen application, on the functional quality of tomato fruits. Chemical assays and untargeted metabolomics were applied to investigate Rhizoglomus irregulare and Funneliformis mosseae administration (both being arbuscular mycorrhiza, AMF), under optimal or low N input conditions, alone or coupled to Trichoderma atroviride application. The coupling of AMF and Trichoderma fungal inoculations resulted in a synergistic biostimulant effect on tomato fruits under sub-optimal fertility, revealing improved concentrations of carotenoid compounds—B-carotene (0.647 ± 0.243 mg/100 g), Z-carotene (0.021 ± 0.021 mg/100 g), 13-z-lycopene (0.145 ± 0.052 mg/100 g) and all-trans-lycopene (12.586 ± 1.511 mg/100 g), and increased values for total phenolic content (12.9 ± 2.9 mgGAE/g), total antioxidant activity (phosphomolybdenum, 0.9 ± 0.2 mmolTE/g), radical scavenging activity (DPPH, 3.4 ± 3.7 mgTE/g), reducing power (FRAP, 23.6 ± 6.3 mgTE/g and CUPRAC, 37.4 ± 7.6 mg TE/g), and enzyme inhibitory activity (AChE, 2.4 ± 0.1 mg GALAE/g), when compared to control. However, evidence of carotenoid and bioactive compounds were exclusively observed under the sub-optimal fertility and no significant differences could be observed between the biostimulant treatment and control under optimal fertility.

Intensive greenhouse vegetable production is often associated with a decline of crop productivity due to the increase of soil salinity and/or a reduction of biological fertility. The aim of the current work was to assess the effects of three organic fertilizers on morpho-physiological and agronomic traits of greenhouse lettuce as well as soil enzyme activity under poor soil quality conditions. The tested organic fertilizers (poultry manure, vinasse-based fertilizer, and insect’s frass fertilizer) were applied pre-planting at the same equivalent nitrogen (N) rate (90 kg N ha−1). Laboratory incubation assay results showed that vinasse-based fertilizer was the most suitable fertilizer in supplying the mineral N in the short term. All fertilizers increased shoot fresh and dry weight compared to unfertilized control with a more pronounced effect (+75%) with vinasse-based fertilizer and insect’s frass. Insect frass reduced by 27% the leaf nitrate concentration in comparison with the other treatments. The toxic heavy metal Pb was 46% lower in all organically fertilized lettuce leaves. Soil enzymatic activities of acid phosphatase, alkaline phosphatase, arylsulfatase (ArS), N-acetyl-β-D-glucosaminidase (NAGase), dehydrogenase, and total hydrolase (THA) were enhanced by poultry manure and insect’s frass in comparison with unfertilized control while vinasse-based fertilizer increased ArS, NAGase, and THA. Taken together, our data demonstrate that the application of organic fertilizers especially vinasse-based fertilizer and insect’s frass during intensive crop production is a suitable approach for mitigating the negative impact of soil salinity, enhancing soil biological fertility, and improving agronomic performance of greenhouse lettuce.

Microbial-based biostimulants can improve crop productivity by modulating cell metabolic pathways including hormonal balance. However, little is known about the microbial-mediated molecular changes causing yield increase. The present study elucidates the metabolomic modulation occurring in pepper (Capsicum annuum L.) leaves at the vegetative and reproductive phenological stages, in response to microbial-based biostimulants. The arbuscular mycorrhizal fungi Rhizoglomus irregularis and Funneliformis mosseae, as well as Trichoderma koningii, were used in this work. The application of endophytic fungi significantly increased total fruit yield by 23.7% compared to that of untreated plants. Multivariate statistics indicated that the biostimulant treatment substantially altered the shape of the metabolic profile of pepper. Compared to the untreated control, the plants treated with microbial biostimulants presented with modified gibberellin, auxin, and cytokinin patterns. The biostimulant treatment also induced secondary metabolism and caused carotenoids, saponins, and phenolic compounds to accumulate in the plants. Differential metabolomic signatures indicated diverse and concerted biochemical responses in the plants following the colonization of their roots by beneficial microorganisms. The above findings demonstrated a clear link between microbial-mediated yield increase and a strong up-regulation of hormonal and secondary metabolic pathways associated with growth stimulation and crop defense to environmental stresses.

The species Pantoea agglomerans includes strains that are agronomically relevant for their growth-promoting or biocontrol traits. Molecular analysis demonstrated that the IPDC pathway involved in the conversion of tryptophan (Trp) to indole-3-acetic acid (IAA) is highly conserved among P. agglomerans strains at both gene and protein levels. Results also indicated that the promoter region controlling the inducible expression of ipdC gene differs from the model system Enterobacter cloacae, which is in accordance with the observation that P. agglomerans accumulates higher levels of IAA when cells are collected in the exponential phase of growth. To assess the potential applications of these microorganisms for IAA production, P. agglomerans C1, an efficient auxin-producer strain, was cultivated in 5 L fermenter so as to evaluate the effect of the medium formulation, the physiological state of the cells, and the induction timing on the volumetric productivity. Results demonstrated that higher IAA levels were obtained by using a saline medium amended with yeast extract and saccharose and by providing Trp, which acts both as a precursor and an inducer, to a culture in the exponential phase of growth. Untargeted metabolomic analysis revealed a significant effect of the carbon source on the exometabolome profile relative to IAA-related compounds and other plant bioactive signaling molecules. The IAA-enriched metabolites secreted in the culture medium by P. agglomerans C1 were used as plant biostimulants to run a series of trials at a large-scale nursery farm. Tests were carried out with in vitro and ex vitro systems following the regular protocols used for large-scale plant tree agamic propagation. Results obtained with 4,540 microcuttings of Prunus rootstock GF/677 and 1,080 plantlets of Corylus avellana L. showed that metabolites from strain C1 improved percentage of rooted-explant, number of adventitious root formation, plant survival, and quality of plant as vigor, with an increase in the leaf area between 17.5 and 42.7% compared to IBA-K (indole-3-butyric acid potassium salt)–treated plants.

Microbial and non-microbial plant biostimulants have been successfully used to improve agriculture productivity in a more sustainable manner. Since the mode of action of biostimulants is still largely unknown, the present work aimed at elucidating the morpho-physiological and metabolomic changes occurring in maize (Zea mays L.) leaves and roots following seed treatment with (i) a consortium of two beneficial fungi [arbuscular mycorrhizal fungi (AMF) and Trichoderma koningii TK7] and rhizobacteria, (ii) a protein hydrolyzate-based biostimulant (PH) alone, or (iii) in combination with a consortium of T. koningii TK7 and rhizobacteria. The application of PH alone or in combination with Trichoderma elicited significant increases (+16.6%) in the shoot biomass compared to untreated maize plants, whereas inoculation with AMF + Trichoderma elicited significant increases in root dry biomass (+48.0%) compared to untreated plants. Distinctive metabolomic signatures were achieved from the different treatments, hence suggesting that different molecular processes were involved in the plants response to the biostimulants. The metabolic reprogramming triggered by the treatments including the protein hydrolyzate was hierarchically more pronounced than the application of microorganisms alone. Most of the differential metabolites could be ascribed to the secondary metabolism, with phenylpropanoids and terpenes being the most represented compounds. The application of PH triggered an accumulation of secondary metabolites, whereas the opposite trend of accumulation was seen in the case of microorganisms alone. The increase in biomass could be related to two processes, namely the modulation of the multilayer phytohormone interaction network and a possible increase in nitrogen use efficiency via the GS-GOGAT system.

In this study, an untargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS) was used for investigating changes in chemical profiles of cow milk considering diets based on mycotoxins-contaminated corn silages. For this purpose, 45 milk samples were classified into five clusters according to the corn silage contamination profile, namely (1) low levels of Aspergillus- and Penicillium-mycotoxins; (2) low levels of fumonisins and other Fusarium-mycotoxins; (3) high levels of Aspergillus-mycotoxins; (4) high levels of non-regulated Fusarium-mycotoxins; (5) high levels of fumonisins and their metabolites, and subsequently analyzed by UHPLC-HRMS followed by a multivariate statistical analysis (both unsupervised and supervised statistical approaches). Overall, the milk metabolomic profile highlighted potential correlations between the quality of contaminated corn silages (as part of the total mixed ration) and milk composition. Metabolomics allowed to identify 628 significant milk metabolites as affected by the five levels of corn silage contamination considered, with amino acids and peptides showing the highest metabolite set enrichment (134 compounds). Additionally, 78 metabolites were selected as the best discriminant of the prediction model built, possessing a variable importance in projection score >1.2. The average Log Fold-Change variations of the discriminant metabolites provided evidence that sphingolipids, together with purine and pyrimidine-derived metabolites were the most affected chemical classes. Also, metabolomics revealed a significant accumulation of oxidized glutathione in milk samples belonging to the silage cluster contaminated by emerging Aspergillus toxins, likely involved in the oxidative imbalance. These preliminary findings provide new insights into the potential role of milk metabolomics to provide chemical indicators of mycotoxins-contaminated corn silage feeding systems.

Bioactive compounds produced by plant growth-promoting bacteria through a fermentation process can be valuable for developing innovative second-generation plant biostimulants. The purpose of this study is to investigate the biotechnological potential of Enterobacter on the production of auxin—a hormone with multiple roles in plant growth and development. The experiments were carried in Erlenmeyer flasks and a 2-L fermenter under batch operating mode. The auxin production by Enterobacter sp. strain P-36 can be doubled by replacing casein with vegetable peptone in the culture medium. Cultivation of strain P36 in the benchtop fermenter indicates that by increasing the inoculum size 2-fold, it is possible to reduce the fermentation time from 72 (shake flask cultivation) to 24 h (bioreactor cultivation) and increase the auxin volumetric productivity from 6.4 to 17.2 mg [IAAequ]/L/h. Finally, an efficient storage procedure to preserve the bacterial auxin was developed. It is noteworthy that by sterilizing the clarified fermentation broth by filtration and storing the filtrated samples at +4 °C, the level of auxin remains unchanged for at least three months.

Salinity in water and soil is one of the major environmental factors limiting the productivity of agronomic and horticultural crops. In basil (Ocimum basilicum L., Lamiaceae) and other Ocimum species, information on the plant response to mild salinity levels, often induced by the irrigation or fertigation systems, is scarce. In the present work, we tested the effectiveness of a microbial-based biostimulant containing two strains of arbuscular mycorrhiza fungi (AMF) and Trichoderma koningii in sustaining greenhouse basil yield traits, subjected to two mild salinity stresses (25 mM [low] and 50 mM [high] modulated by augmenting the fertigation osmotic potential with NaCl) compared to a non-stressed control. The impact of salinity stress was further appraised in terms of plant physiology, morphological ontogenesis and composition in polyphenols and volatile organic compounds (VOC). As expected, increasing the salinity of the solution strongly depressed the plant yield, nutrient uptake and concentration, reduced photosynthetic activity and leaf water potential, increased the Na and Cl and induced the accumulation of polyphenols. In addition, it decreased the concentration of Eucalyptol and β-Linalool, two of its main essential oil constituents. Irrespective of the salinity stress level, the multispecies inoculum strongly benefited plant growth, leaf number and area, and the accumulation of Ca, Mg, B, p-coumaric and chicoric acids, while it reduced nitrate and Cl concentrations in the shoots and affected the concentration of some minor VOC constituents. The benefits derived from the inoculum in term of yield and quality harnessed different mechanisms depending on the degree of stress. under low-stress conditions, the inoculum directly stimulated the photosynthetic activity after an increase of the Fe and Mn availability for the plants and induced the accumulation of caffeic and rosmarinic acids. under high stress conditions, the inoculum mostly acted directly on the sequestration of Na and the increase of P availability for the plant, moreover it stimulated the accumulation of polyphenols, especially of ferulic and chicoric acids and quercetin-rutinoside in the shoots. Notably, the inoculum did not affect the VOC composition, thus suggesting that its activity did not interact with the essential oil biosynthesis. These results clearly indicate that beneficial inocula constitute a valuable tool for sustaining yield and improving or sustaining quality under suboptimal water quality conditions imposing low salinity stress on horticultural crops.

Plant-derived protein hydrolysates (PHs) are an important category of biostimulants able to increase plant growth and crop yield especially under environmental stress conditions. PHs can be applied as foliar spray or soil drench. Foliar spray is generally applied to achieve a relatively short-term response, whereas soil drench is used when a long-term effect is desired. The aim of the study was to elucidate the biostimulant action of PH application method (foliar spray or substrate drench) on morpho-physiological traits and metabolic profile of tomato grown under limited water availability. An untreated control was also included. A high-throughput image-based phenotyping (HTP) approach was used to non-destructively monitor the crop response under limited water availability (40% of container capacity) in a controlled environment. Moreover, metabolic profile of leaves was determined at the end of the trial. Dry biomass of shoots at the end of the trial was significantly correlated with number of green pixels (R2 = 0.90) and projected shoot area, respectively. Both drench and foliar treatments had a positive impact on the digital biomass compared to control while the photosynthetic performance of the plants was slightly influenced by treatments. Overall drench application under limited water availability more positively influenced biomass accumulation and metabolic profile than foliar application. Significantly higher transpiration use efficiency was observed with PH-drench applications indicating better stomatal conductance. The mass-spectrometry based metabolomic analysis allowed the identification of distinct biochemical signatures in PH-treated plants. Metabolomic changes involved a wide and organized range of biochemical processes that included, among others, phytohormones (notably a decrease in cytokinins and an accumulation of salicylates) and lipids (including membrane lipids, sterols, and terpenes). From a general perspective, treated tomato plants exhibited an improved tolerance to reactive oxygen species (ROS)-mediated oxidative imbalance. Such capability to cope with oxidative stress might have resulted from a coordinated action of signaling compounds (salicylic acid and hydroxycinnamic amides), radical scavengers such as carotenoids and prenyl quinones, as well as a reduced biosynthesis of tetrapyrrole coproporphyrins.

Designing and developing new biostimulants is a crucial process which requires an accurate testing of the product effects on the morpho-physiological traits of plants and a deep understanding of the mechanism of action of selected products. Product screening approaches using omics technologies have been found to be more efficient and cost effective in finding new biostimulant substances. A screening protocol based on the use of high-throughput phenotyping platform for screening new vegetal-derived protein hydrolysates (PHs) for biostimulant activity followed by a metabolomic analysis to elucidate the mechanism of the most active PHs has been applied on tomato crop. Eight PHs (A–G, I) derived from enzymatic hydrolysis of seed proteins of Leguminosae and Brassicaceae species were foliarly sprayed twice during the trial. A non-ionic surfactant Triton X-100 at 0.1% was also added to the solutions before spraying. A control treatment foliarly sprayed with distilled water containing 0.1% Triton X-100 was also included. Untreated and PH-treated tomato plants were monitored regularly using high-throughput non-invasive imaging technologies. The phenotyping approach we used is based on automated integrative analysis of photosynthetic performance, growth analysis, and color index analysis. The digital biomass of the plants sprayed with PH was generally increased. In particular, the relative growth rate and the growth performance were significantly improved by PHs A and I, respectively, compared to the untreated control plants. Kinetic chlorophyll fluorescence imaging did not allow to differentiate the photosynthetic performance of treated and untreated plants. Finally, MS-based untargeted metabolomics analysis was performed in order to characterize the functional mechanisms of selected PHs. The treatment modulated the multi-layer regulation process that involved the ethylene precursor and polyamines and affected the ROS-mediated signaling pathways. Although further investigation is needed to strengthen our findings, metabolomic data suggest that treated plants experienced a metabolic reprogramming following the application of the tested biostimulants. Nonetheless, our experimental data highlight the potential for combined use of high-throughput phenotyping and metabolomics to facilitate the screening of new substances with biostimulant properties and to provide a morpho-physiological and metabolomic gateway to the mechanisms underlying PHs action on plants.

The implementation of sustainable agriculture encompasses practices enhancing the activity of beneficial soil microorganisms, able to modulate biogeochemical soil cycles and to affect soil fertility. Among them, arbuscular mycorrhizal fungi (AMF) establish symbioses with the roots of most food crops and play a key role in nutrient uptake and plant protection from biotic and abiotic stresses. Such beneficial services, encompassing improved crop performances, and soil resources availability, are the outcome of the synergistic action of AMF and the vast communities of mycorrhizospheric bacteria living strictly associated with their mycelium and spores, most of which showing plant growth promoting (PGP) activities, such as the ability to solubilize phosphate and produce siderophores and indole acetic acid (IAA). One of the strategies devised to exploit AMF benefits is represented by the inoculation of selected isolates, either as single species or in a mixture. Here, for the first time, the microbiota associated with a commercial AMF inoculum was identified and characterized, using a polyphasic approach, i.e., a combination of culture-dependent analyses and metagenomic sequencing. Overall, 276 bacterial genera were identified by Illumina high-throughput sequencing, belonging to 165 families, 107 orders, and 23 phyla, mostly represented by Proteobacteria and Bacteroidetes. The commercial inoculum harbored a rich culturable heterotrophic bacterial community, whose populations ranged from 2.5 to 6.1 × 106 CFU/mL. The isolation of functional groups allowed the selection of 36 bacterial strains showing PGP activities. Among them, 14 strains showed strong IAA and/or siderophores production and were affiliated with Actinomycetales (Microbacterium trichotecenolyticum, Streptomyces deccanensis/scabiei), Bacillales (Bacillus litoralis, Bacillus megaterium), Enterobacteriales (Enterobacter), Rhizobiales (Rhizobium radiobacter). This work demonstrates for the first time that an AMF inoculum, obtained following industrial production processes, is home of a large and diverse community of bacteria with important functional PGP traits, possibly acting in synergy with AMF and providing additional services and benefits. Such bacteria, available in pure culture, could be utilized, individually and/or in multispecies consortia with AMF, as biofertilizers and bioenhancers in sustainable agroecosystems, aimed at minimizing the use of chemical fertilizers and pesticides, promoting primary production, and maintaining soil health and fertility.

Sweet cherry is a high value crop and the economic success of its cultivation depends not only on yield but also on fruit visual and nutritional quality attributes that influence consumer acceptability, as well as on fruit post-harvest performance and resistance to cracking. During the last few decades, cherry growers have tried to achieve these goals through exogenous applications of synthetic plant hormones and/or nutrients, but there is growing concern about the sustainability of the extensive use of these compounds in agriculture. For this reason, there is increasing interest in the possible adoption of different classes of biostimulants as sustainable alternatives to plant growth regulators. This research aimed to study the impact of foliar application of a novel tropical-plant extract, performed between full bloom and fruit set, on the yield and fruit quality of two important commercial sweet cherry cultivars, Kordia and Regina. The experimental design included a commercial control involving the application of a cytokinin promoter. In both cultivars, the tropical-plant extract induced significant increases in fruit yield. In addition, in the cultivar Kordia, the tropical-plant extract enhanced fruit calcium concentration, soluble solids content, flesh firmness, and skin color by 26.2%, 11.8%, 6.7%, and 12.0% (of fruits with mahogany skin color), respectively. Our results suggest that the tropical-plant extract tested as a biostimulant may be a sustainable and effective alternative to the exogenous application of synthetic hormones for sweet cherry cultivation.

The city of Apollonia, located to the South-west of modern Albania, was founded in 588 B.C. by Corinthian and Corcyrian people, and was later described by Cicero as an admirabilis urbs. In 44 B.C. when Augustus and Agrippa were studying rhetoric in Apollonia, they got to know of the killing of Caesar and were helped by the local people to return to Rome and take the power back. The city, located on the right bank of the river Aoos, only 12 km from the sea, was a crossroads and a filter for different cultures: Illyrians, Macedonians, the Greeks and the Romans have inhabited the territory over the centuries. The domus, dated between 2nd and 3rd century A.D., represents the connection of the “residential fashion” present at the East and West of the Adriatic Sea. The domus is known in the literature as the “Athena house” because of the statue discovered by the archaeologists inside of it, but also as the “D house” because it was unearthed in “Sector D” of the excavation. The first goal of the research is to analyse the architectural and decorative aspects of the house to understand the tastes of this site. The second goal is to understand the needs of the customer’s social and economic selfrepresentation and the skills of the craftsmen who worked there.

A greenhouse experiment was established with loquat plants to investigate the role of arbuscular mycorrhizal fungi (AMF) in the control of the white root rot fungus Armillaria mellea and to determine the changes produced in the plant metabolome. Plants inoculated with two AMF, Rhizoglomus irregulare and a native AMF isolate from loquat soils, were infected with Armillaria. Although mycorrhization failed to control the Armillaria root infection, the increased growth of infected plants following inoculation with the native mycorrhizal isolate suggests an initial tolerance towards Armillaria. Overall, metabolomics allowed highlighting the molecular basis of the improved plant growth in the presence of Armillaria following AMF colonization. In this regard, a wide and diverse metabolic response was involved in the initial tolerance to the pathogen. The AMF-mediated elicitation altered the hormone balance and modulated the production of reactive oxygen species (mainly via the reduction of chlorophyll intermediates), possibly interfering with the reactive oxygen species (ROS) signaling cascade. A complex modulation of fucose, ADP-glucose and UDP-glucose, as well as the down-accumulation of lipids and fatty acids, were observed in Armillaria-infected plants following AMF colonization. Nonetheless, secondary metabolites directly involved in plant defense, such as DIMBOA and conjugated isoflavone phytoalexins, were also involved in the AMF-mediated plant response to infection.

Plant-derived protein hydrolysates (PHs) have received increased attention in the last decade because of their potential to improve yield, nutritional quality as well as tolerance to abiotic stressors. The current study investigated the effects and the molecular mechanisms of a legume-derived PH under optimal and sub-optimal nitrogen (N) concentrations (112 and 7 mg L−1, respectively) in tomato (Solanum lycopersicum L.). Growth and mineral composition of tomato plants treated with PHs by foliar spray or substrate drench were compared to untreated plants. In addition, the expression was determined of genes encoding ammonium and nitrate transporters and seven enzymes involved in N metabolism: nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase 1 (GS1), glutamine synthetase 2 (GS2), ferredoxin-dependent glutamate synthase (GLT), NADH-dependent glutamate synthase (GLS), and glutamate dehydrogenase (GDH). The root and total plant dry weight, SPAD index and leaf nitrogen content were higher by 21, 17, 7, and 6%, respectively, in plants treated by a substrate drench in comparison to untreated tomato plants, whereas foliar application of PH gave intermediate values. PH-treated plants grown with lower N availability showed reduced expression of NR and NiR as well as of nitrate and ammonium transporter transcripts in both leaf and root tissues in comparison with untreated plants; this was especially pronounced after application of PH by substrate drench. Conversely, the transcript level of an amino acid transporter gene was up-regulated in comparison with untreated plants. At high N regime, the transcript levels of the ammonium and amino acid transporters and also NR, NiR, and GLT were significantly up-regulated in root after PH foliar and substrate drench applications compared with untreated plants. An up-regulation was also observed for GS1, GS2, and GDH transcripts in leaf after substrate drench. These results highlighted the potential benefits of using legume PH in vegetable production systems to increase growth and N-nutritional status of plants.

Plant biostimulants are receiving great interest for boosting root growth during the first phenological stages of vegetable crops. The present study aimed at elucidating the morphological, physiological, and metabolomic changes occurring in greenhouse melon treated with the biopolymer-based biostimulant Quik-link, containing lateral root promoting peptides, and lignosulphonates. The vegetal-based biopolymer was applied at five rates (0, 0.06, 0.12, 0.24, or 0.48 mL plant-1) as substrate drench. The application of biopolymer-based biostimulant at 0.12 and 0.24 mL plant-1 enhanced dry weight of melon leaves and total biomass by 30.5 and 27.7%, respectively, compared to biopolymer applications at 0.06 mL plant-1 and untreated plants. The root dry biomass, total root length, and surface in biostimulant-treated plants were significantly higher at 0.24 mL plant-1 and to a lesser extent at 0.12 and 0.48 mL plant-1, in comparison to 0.06 mL plant-1 and untreated melon plants. A convoluted biochemical response to the biostimulant treatment was highlighted through UHPLC/QTOF-MS metabolomics, in which brassinosteroids and their interaction with other hormones appeared to play a pivotal role. Root metabolic profile was more markedly altered than leaves, following application of the biopolymer-based biostimulant. Brassinosteroids triggered in roots could have been involved in changes of root development observed after biostimulant application. These hormones, once transported to shoots, could have caused an hormonal imbalance. Indeed, the involvement of abscisic acid, cytokinins, and gibberellin related compounds was observed in leaves following root application of the biopolymer-based biostimulant. Nonetheless, the treatment triggered an accumulation of several metabolites involved in defense mechanisms against biotic and abiotic stresses, such as flavonoids, carotenoids, and glucosinolates, thus potentially improving resistance toward plant stresses.

Plant-derived protein hydrolysates (PHs) have gained prominence as plant biostimulants because of their potential to increase the germination, productivity and quality of a wide range of horticultural and agronomic crops. Application of PHs can also alleviate the negative effects of abiotic plant stress due to salinity, drought and heavy metals. Recent studies aimed at uncovering the mechanisms regulating these beneficial effects indicate that PHs could be directly affecting plants by stimulating carbon and nitrogen metabolism, and interfering with hormonal activity. Indirect effects could also play a role as PHs could enhance nutrient availability in plant growth substrates, and increase nutrient uptake and nutrient-use efficiency in plants. Moreover, the beneficial effects of PHs also could be due to the stimulation of plant microbiomes. Plants are colonized by an abundant and diverse assortment of microbial taxa that can help plants acquire nutrients and water and withstand biotic and abiotic stress. The substrates provided by PHs, such as amino acids, could provide an ideal food source for these plant-associated microbes. Indeed, recent studies have provided evidence that plant microbiomes are modified by the application of PHs, supporting the hypothesis that PHs might be acting, at least in part, via changes in the composition and activity of these microbial communities. Application of PHs has great potential to meet the twin challenges of a feeding a growing population while minimizing agriculture’s impact on human health and the environment. However, to fully realize the potential of PHs, further studies are required to shed light on the mechanisms conferring the beneficial effects of these products, as well as identify product formulations and application methods that optimize benefits under a range of agro-ecological conditions.

Scientific investigations are being increasingly devoted to biostimulant effects on vegetable yield and quality, with the perspective of sustainable crop management. Two farming systems (conventional or organic) in factorial combination with two biostimulant treatments (tropical plant extract (PE); legume-derived protein hydrolysate (PH)) plus a non-treated control were compared in terms of tomato fruit yield, yield components, mineral composition, functional and nutritional indicators. PE- and PH-based biostimulants resulted in higher plant biomass, PH even in higher leaf area index, compared to non-treated control. Marketable yield was not significantly affected by farming system. PH and PE gave higher yield than non-treated control. PH treatment led to higher fruit number than the control, whereas PE incurred significant increase in yield only under organic farming. The mean fruit weight attained the highest value upon PE application under conventional management. Colour component a* (redness) was higher with the conventional system compared to the organic one, whereas an opposite trend was shown by the organic acids malate, oxalate and isocitrate. Irrespective of the farming system, the soluble solids, fruit brightness (L*) and redness as well as the target organic acids malate, oxalate, citrate and isocitrate were significantly higher than untreated plants by 10.1%, 16.1%, 19.8%, 18.9%, 12.1%, 13.5% and 26.8%, respectively, with no significant differences between the PH- and PE-based biostimulants. Higher lipophilic activity and total ascorbic acid concentration but lower lycopene were recorded under organic management. PE and PH application resulted in higher total phenol and ascorbic acid as well as in lycopene content, and lipophilic antioxidant activity than the non-treated control. Biostimulants proved to be an effective sustainable tool for enhancing tomato fruit yield and functional quality both under conventional and organic vegetable systems.

The emerging role of plant biostimulants in enhancing nutrient efficiency is important for maintaining soil fertility under sub-optimal nutrient regimens. We aimed to elucidate the morpho-physiological and biochemical effects as well as mineral composition changes of greenhouse jute (Corchorus olitorius L.) treated with a commercial vegetal-derived biostimulant from a tropical plant extract (PE; Auxym®, Italpollina, Rivoli Veronese, Italy). Plants were sprayed in weekly intervals with a solution containing 2 mL·L−1 PE. Jute plants were supplied with three nutrient solution concentrations: full-, half-, and quarter-strength. Decreasing macronutrient concentrations in the nutrient solution (NS), especially at quarter-strength, triggered a decrease in several morphological (plant height, leaf number, and dry biomass) and physiological (net CO2 assimilation rate (ACO2) and SPAD (Soil Plant Analysis Development) index) parameters. PE application triggered specific ameliorative effects in terms of fresh yield at both half- and quarter-strength nutrient solution (15.5% and 29.5%, respectively). This was associated with an enhancement in ACO2, SPAD index, and especially the nutritional status (high nitrate, K, and Mg contents, and low Na content). The foliar application of PE, strongly increased chlorophyll b content, enhancing jute plant adaptation to fluctuating light and therefore the efficiency of photosynthesis, positively affecting starch, soluble proteins, and total amino acids content but only when jute plants were irrigated with full-strength NS, compared to the respective control treatment. At lower nutrient strength, PE reprogrammed the nitrogen distribution, allowing its remobilization from glutamate, which was quantitatively the major amino acid under lower nutrient strength, but not from chlorophylls, thus maintaining efficient photosynthesis. We confirmed that PE Auxym® acts in a balanced manner on the main metabolic pathways of the plant, regulating the uptake and transport of mineral nutrients and protein synthesis, increasing the accumulation of essential amino acids under full nutritive solutions, and re-distributing nitrogen from amino acids to allow leaf growth and expansion even under sub-optimal nutrient conditions. Overall, the use of natural plant biostimulants may be a potential solution in low-input conditions, where environmental constraints and restricted use of fertilizers may affect potential crop productivity.

Plant biostimulants (PBs) such as protein hydrolysates and seaweed extracts are attracting the increasing interest of scientists and vegetable growers for their potential toenhance yield and nutritional quality. The current study assessed crop productivity, leaf colorimetry, mineral profile and bioactive compounds of greenhouse spinach in response to the foliar application of three PBs: legume-derived protein hydrolysate [PH], extract of seaweed Ecklonia maxima or mixture of vegetal oils, herbal and seaweed Ascophyllum nodosum extracts. Plants were PB-treated at a rate of 3 mL L−1 four times during their growth cycle at weekly intervals. Foliar PB applications enhanced fresh yield, dry biomass and leaf area of spinach in comparison with untreated plants. Improved yield performance with PB applications was associated with improved chlorophyll biosynthesis (higher SPAD index). The three PB treatments elicited an increase in bioactive compounds (total phenols and ascorbic acid), thus raised the functional quality of spinach. The application of PH enhanced K and Mg concentrations and did not result in increased nitrate accumulation as observed with the other two PB treatments. Our findings can assist vegetable farmers and the agro-food industry in adopting innovative and sustainable tools such as PB for complementing a high yield with premium quality.